service d ’ h é matologie on behalf of the « western algerian group of bone marrow transplant »...

Service d’Hématologie

On behalf of the « Western Algerian Group of Bone Marrow Transplant »WAG-BMT

Haematology and Cell Therapy DepatmentHemobiology Department

Viability of non cryopreserved CD34+Stem Cells

Service d’HématologieIntroduction

Autologous stem cell transplantation (ASCT) after a high dose conditioning is an established treatment modality with definitive indications for many hematological disorders.

However, this line of treatment requires many expensive resources, such as freezing of the harvest product in order to maintain cell viability until stem-cell reinfusion and the use of growth factors for the management of neutropenia.

Service d’Hématologie Aim of the study

To study the viability of CD34+ stem cells stored at +4°C

To demonstrate the feasibility and the safety of the ASCT without any cryopreservation neither the use of growth factors.

Service d’Hématologie Study design STEP 1: Short Conditioning regiments

ASCT in MM (MEL200) in 75 patientsASCT in HL (CBV) in 17 patients

STEP 2: Long conditioning regimentsASCT in HL (BEAM) in 02 patientsASCT in HL and NHL (EAM) in 15 patients


Figure 1. Schema of procedure.

STEP 1Material and methods

75 patients with Multiple Myeloma


17 patients with Hodgkin Lymphoma



Wannesson L et al. Annals of Oncology 18: 623–632, 2007




A 2ml sample was taken from 10 collection bags and then was sent to the flow cytometry laboratory where it was stored in the refrigerator at +4°C.

A CD34+ stem cells count were performed using a “Lyse no wash double platform CD34+ flow cytometry assay”

The viability of CD34+ cells was assessed with 7'AAD immediately after the end of apherisis (H0) then at (H24), (H48) and finally at (H72).


STEP 1Results

H0 H24 H48 H72Viability (median) 98% 97.6% 96% 95%

Viability (range) 97.4 – 100 92.4 – 99.4 91.6-98 91-96


STEP 1Results

y = -1,06x + 99,3R2 = 0,9571

p=0.05

CD34+ stem cells viability kenetic curve


STEP 1Results

Brahimi M et al. Revue Algerienne D’hematologie 2011 ; 5 :46-48.


STEP 1Results

Time of storage of the Harvest bags at +4°C

n=200


STEP 1Results

CBV (HODGKIN)

(n = 17) Mel200 (MYELOMA)

(n=75)CD34+ cells/Kg 3,66 (2,3-13,22) 3,66 (1,5 - 9,7)Duration of neutropenia 13(9-24) 10(06-17)Duration of thrombocytopenia, 12(9-37) 13(9-24)No. of platelet transfusions, 1(0-2) 1(0-3)No. of red blood cell transfusions, 2(0-8) 2(0-9)Duration of intravenous antibiotics, 9(0-15) 4(0-13)Duration of hospitalization, 21(15-37) 17(12-30)Transplant Related Mortality 01 00

Service d’Hématologie Mel200 protocol

Bekadja MA et al. Hematol Oncol Stem Cell Ther 2012; 5(1): 49-53

Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012.page 140 (Abstract 905)


Bekadja MA et al. Revue Algerienne D’hematologie 2011 ; 5 :50-53.

CBV protocol


STEP 2«BEAM» VS «-EAM»

BEAM (n=2) -EAM (n=15)

BCNU (mg/m2) 300 (D-2) ---

VP16 (mg/m2) 200 (D-5 to D-2) 200 (D-5 to D-2)

ARAC (mg/m2) 400 (D-5 to D-2) 2000 (D-5 to D-2)

MELPHALAN (mg/m2) 140 (D-1) 140 (D-1)


STEP2Material and methods

15 Patients with Hodgkin Lymphoma

Talhi S et al. Hématologie, Vol. 1, Supplement 1, Mars 2012: pp 213 (Abstract)

Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012.page 162 (Abstract 1140)


STEP2RESULTS

n=30

Time of storage of the Harvest bags


STEP2RESULTS

BEAM (HODGKIN)

(n = 2) -EAM (HODGKIN)

(n=15)CD34+ cells/Kg 6,98 (4,42-9,53) 3,68 (2,71 - 6,15)Duration of neutropenia 12(11-13) 13(10-32)Duration of thrombocytopenia, 11(11-12) 13(9-24)No. of platelet transfusions, 2(1-4) 3(2-4)No. of red blood cell transfusions 2(2-3) 4(2-9)Transplant Related Mortality 00 00


DAY : 0 (n=10) DAY : 6 (n=10)Viability ( % )Per Bag 93,77 ( 70-100 ) 82,18 ( 54,24 – 94 )

MNC ( % )Per Bag 51,24 ( 25,85 – 71,21 ) 57,05 ( 39,71 – 75 )

CD34 (% )Per Bag 0,73 ( 0,16 – 1,94 ) 0,34 ( 0,19 – 0,54 )

STEP2RESULTS

Brahimi M et al. Unpublished data.

Service d’Hématologie Conclusion

We conclude that high-dose chemotherapy with non-frozen peripheral stem cells is safe in terms of haematopoietic reconstitution even without using growth factors.


Department of HemobiologyEHU 1st November, Oran, Algeria


23

Department of Hematology and Stem Cell Therapy EHU 1st November Oran, Algeria


Professor Mohamed Amine BEKADJA

Bekadja MA et al. Hematol Oncol Stem Cell Ther 2012; 5(1): 49-53

Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012., (Abstract 905)

Bekadja MA et al. Revue Algérienne d’Hématologie 2011 ; 5 :50-53.

Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012, (Abstract 1140)

Chief of Biology Pôle in EHU 1st November

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